Molecular identification and quantification of bacteria from endodontic infections using real‐time polymerase chain reaction

Blome, B.; Braun, Andreas; Sobarzo, V.; Jepsen, Søren

doi:10.1111/j.1399-302x.2008.00440.x

Cited by 90 publications

(104 citation statements)

References 40 publications

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“…6,7 However, in the present study intracanal dressing was not able to eliminate or reduce the target species investigated and did not reduce DNA concentration when compared with the initial sample. The present results are in clear agreement with those of Blome et al 25 They observed that a Ca(OH) 2 intracanal dressing placed for 14 days failed to achieve a further reduction of total bacterial counts when compared to the values observed immediately after chemo-mechanical preparation. 25 Moreover, this is also in agreement with the findings of Sakamoto et al, 17 …”

Section: Discussionsupporting

confidence: 83%

“…The present results are in clear agreement with those of Blome et al 25 They observed that a Ca(OH) 2 intracanal dressing placed for 14 days failed to achieve a further reduction of total bacterial counts when compared to the values observed immediately after chemo-mechanical preparation. 25 Moreover, this is also in agreement with the findings of Sakamoto et al, 17 …”

Section: Discussionsupporting

confidence: 83%

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PCR identiﬁcation of endodontic pathogens and DNA quantiﬁcation in samples from teeth with posttreatment apical periodontitis

et al. 2014

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The aim of this clinical study was to quantify the concentration of DNA and to detect selected bacterial species from samples of infected root-fi lled teeth with post-treatment apical periodontitis after removal of gutta-percha (S1), after chemo-mechanical preparation at the fi rst appointment (S2), 5 days after the canal was fi lled with sterile physiological solution (S3), after reinstrumentation at the second appointment (S4), and 14 days after an intracanal dressing was placed at the third appointment (S5). Methods: Fifteen root-fi lled teeth were selected. Removal of gutta-percha was performed using the crown-down technique. Chemo-mechanical preparation was performed with hand fi les associated with 2% chlorhexidine gel. An intracanal dressing based on Ca(OH) 2 was used. DNA was extracted from the samples and 14 endodontic 16S rDNA species-specifi c primers were tested. The concentration of DNA was quantifi ed using a NanoDropTM 2000 spectrophotometer. Results: Bacteria were present in all cases at all sampling times, as revealed by a universal primer. DNA was isolated from all samples, with an average concentration of 4.24 ± 2.9 ng/µL (S1), 3.39 ± 1.54 ng/µL (S2), 4.0 ± 1.94 ng/µL (S3), 2.66 ± 0.98 ng/µL (S4) and 3.97 ± 2.32 ng/µL (S5). Parvimonas micra and Enterococcus faecalis (S1), P. micra (S2), Porphyromonas endodontalis and E. faecalis (S3), E. faecalis and Prevotella nigrescens (S4/S5) were the species most frequently deteced. DNA concentration reductions were detected between S3 and S4 (p = 0.0256), whereas an increase was found between S4 and S5. Conclusion: A wide variety of bacterial species was detected in root-fi lled teeth with post-treatment apical periodontitis. Moreover, the use of an intracanal dressing was unable to further reduce the concentration of bacterial DNA.descrIPtors | Endodontics; Treatment Failure; Periapical Periodontitis; Bacteria; Polymerase Chain Reaction; DNA; Chlorhexidine.resuMo | Identifi cação de patógenos endodônticos por PCR e quantifi cação de DNA em amostras extraídas de dentes com periodontite apical pós-tratamento • Objetivo: O objetivo deste estudo clínico foi quantifi car a concentração de DNA e detectar algumas espécies bacterianas de amostras de dentes tratados endodonticamente com periodontite apical após a remoção da guta-percha (S1), após o preparo químico-mecânico na primeira sessão (S2), 5 dias após o preenchimento do canal com solução fi siológica estéril (S3), após reinstrumentação na segunda sessão (S4), e 14 dias após a inserção da medicação intracanal na terceira sessão (S5). Métodos: Quinze dentes tratados endodonticamente foram selecionados. A remoção da guta-percha foi realizada por meio da técnica coroa-ápice. Utilizaram-se limas manuais associadas à clorexidina gel a 2% durante o preparo químico-mecânico. A medicação intracanal selecionada foi à base de hidróxido de cálcio. DNA foi isolado das amostras e foram investigadas 14 espécies bacterianas (primer espécie-específi co16S rDNA). A concentração de DNA foi quantifi cada utilizando o espect...

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Section: Discussionsupporting

confidence: 83%

Section: Discussionsupporting

confidence: 83%

PCR identiﬁcation of endodontic pathogens and DNA quantiﬁcation in samples from teeth with posttreatment apical periodontitis

et al. 2014

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“…In cases of primary endodontic infections, there is a higher bacterial load than in secondary infections [6][7] the bacterial load is reduced by at least 95% and the remaining viable bacteria may be the cause of refractory lesions 8 . Teeth with endodontic failure have been associated with the presence of Enterococcus species [9][10] .…”

Section: Introductionmentioning

confidence: 99%

Enterococcus spp. isolated from root canals with persistent chronic apical periodontitis in a Chilean population

Sánchez‐Sanhueza¹,

González-Rocha²,

Dominguez³

et al. 2015

Braz. J. Oral Sci.

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Aim: To isolate and identify in a Chilean population, Enterococcus spp. from root canals with persistent chronic apical periodontitis (CAP) and to investigate the potential correlation between the bacteria and the observed clinical features. Methods: Twenty patients with indication for endodontic retreatment due to persistent CAP were selected. Data from patient general health and dental clinical history were recorded. During retreatment, a microbial sample was obtained from the root canal and inoculated in a selective Enterococcus medium. Using bacterial cultivation methods, bacterial isolates belonging to the genus Enterococcus were identified. The relationship between the number of colony-forming units of Enterococcus spp. and patient clinical data was assessed statistically by the Pearson Chi square and Fisher exact tests. Finally, a Polymerase Chain Reaction (PCR) assay to determine the most prevalent species of Enterococcus spp. was conducted in the clinical samples, and the results were analyzed by a proportion comparison test. Results: Enterococcus spp. strains were isolated in 70% of the patients. Most of them (98.8%) accounted for Enterococcus faecalis and only 1.2% for Enterococcus faecium. A high frequency of E. faecalis was found in teeth with inadequate endodontic treatment or dental crown restorations. Conclusions: This study concluded that E. faecalis is prevalent in root canals with persistent CAP in a Chilean population. E. faecium as found in a single case with the poorest root canal filling. Further studies are still required to investigate the presence of other species, which may be linked to persistent chronic apical periodontitis.

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“…Os níveis totais antes do preparo, obtidos nesta pesquisa, estão de acordo com aqueles que podemos observar nos trabalhos já publicados, que variam entre 10 6 e 10 7 em média, mas podendo apresentar limites mínimos e máximos de 10 3 até 10 8 . A média aqui encontrada foi ligeiramente maior (10 8 ) (Vianna et al, 2006;Sakamoto et al, 2007;Blome et al, 2008 (Tang et al, 2004;Vianna et al, 2006;Sakamoto et al, 2007;Blome et al, 2008;Vianna et al, 2008;Jiang et al, 2009;Roças;Siqueira 2010;Alves et al, 2012).…”

Section: Resultsunclassified

“…Os trabalhos mais recentes, que avaliaram a capacidade do preparo químico mecânico na redução da microbiota endodôntica, apontam que a redução gira em torno de 95 a 99% em média (Tang et al, 2004;Vianna et al, 2006;Sakamoto et al, 2007;Blome et al, 2008;Vianna et al, 2008;Jiang et al, 2009;Roças;Siqueira 2010;Alves et al, 2012).…”

Section: Preparo Químico Mecânico Do Sistema De Canais Radicularesunclassified

Avaliação in vivo da redução microbiana após preparo do canal radicular com auxílio do sistema EndoVac

Bitencourt¹

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"O conhecimento científico é cumulativo, dependente do desenvolvimento de novas tecnologias e idéias. Nossas verdades permanecem verdadeiras até que alguém demonstre que elas são incompletas. No mínimo, esta perspectiva deveria nos ensinar a defender nossas verdades temporárias com a humildade de quem aprende com o passado." The present study aimed to analyze in vivo the effectiveness of root canal preparation on bacterial reduction in patients's teeth with primary apical periodontitis. Marcelo Gleiser RESUMOSamples were collected from 20 patients before (S1) and after (S2) preparation with ProTaper rotary files, varying only the technique used for irrigation and aspiration:Group A (conventional irrigation -n = 10) and Group B (with the aid of EndoVac system -n = 10). After extraction, the DNA present in the samples was quantified by real-time PCR with the SYBR Green method, identifying the number of 16SrRNA gene copies. In all samples, except for a post-preparation case with EndoVac, copies of the target gene were identified. Average for all cases was 1,6 X 10 8 and 8,8 X 10 5 copies of the 16SrRNA, before and after preparation, respectively. For the groups separately, the same values were 2,0 X 10 8 and 5,5 X 10 5 (conventional) and 1,1 X 10 8 and 1,2 X 10 6 (EndoVac). The mean percentage of reduction was 97.52% (97.02% for the conventional and 98.04% for the EndoVac). The Mann-Whitney test concluded that both techniques significantly reduced the microrganisms after preparation (p<0,0001), with no differences between them (p=0,9705). None of the techniques were effective in the complete elimination of bacteria under this methodology. The antibacterial efficacy of the EndoVac system under this methodology was similar to that obtained with conventional irrigation and aspiration.

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Molecular identification and quantification of bacteria from endodontic infections using real‐time polymerase chain reaction

Abstract: Root canals with primary infections contained a higher bacterial load. Chemo-mechanical root canal preparation reduced bacterial counts by at least 95%.

Cited by 90 publications

References 40 publications

PCR identiﬁcation of endodontic pathogens and DNA quantiﬁcation in samples from teeth with posttreatment apical periodontitis

PCR identiﬁcation of endodontic pathogens and DNA quantiﬁcation in samples from teeth with posttreatment apical periodontitis

Enterococcus spp. isolated from root canals with persistent chronic apical periodontitis in a Chilean population

Avaliação in vivo da redução microbiana após preparo do canal radicular com auxílio do sistema EndoVac

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